CN106718875A - Method for growing dendrobium seedlings - Google Patents
Method for growing dendrobium seedlings Download PDFInfo
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- CN106718875A CN106718875A CN201611027267.5A CN201611027267A CN106718875A CN 106718875 A CN106718875 A CN 106718875A CN 201611027267 A CN201611027267 A CN 201611027267A CN 106718875 A CN106718875 A CN 106718875A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Abstract
The present invention discloses a kind of method for growing dendrobium seedlings, and the method includes：A, the stem of noble dendrobium stem section of belt segment is cultivated in the medium, induce axillary bud；B, when axillary bud growth is highly more than 1cm, by bud point excision, induce Multiple Buds；C, when the growth of all living creatures' bud reaches 1cm~2cm, cut Multiple Buds, Multiple Buds are rised in value and culture of rootage, and will cuts the former stem section continuation of Multiple Buds and cultivated in the medium, evoked callus；D, after Callus formation, callus is cut and is transferred in culture medium and continues to cultivate, induce Multiple Buds；E, Multiple Buds are transferred to Multiplying culture in culture medium；F, when Multiple Buds growing height reaches 2cm~3cm, being transferred in culture medium carries out culture of rootage.Technical scheme can effectively increase the numerous quantity of expansion, so as to produce the stem of noble dendrobium seedling of a large amount of high-quality.
Description
Technical field
The present invention relates to technical field of tissue culture, more particularly to a kind of method for growing dendrobium seedlings.
Background technology
The stem of noble dendrobium is a big quasi-tradition rare traditional Chinese medicine in China, traditionally can be both used for medicinal purpose, and health products can be used as again, together
When, start also to be used as a kind of high-grade soup stock (food) always till now from way back, traditional medicinal of the current stem of noble dendrobium
The height for having obtained people with healthcare function is accepted and liked.
Need to be sprouted with mycosymbiosis offer nutrition under field conditions (factors) due to the stem of noble dendrobium, and germination rate is extremely low, it is wild
Stem of noble dendrobium happiness is shady and cool, moistening, foggy climatic environment of divulging information, extremely harsh to growing environment, meets high light, heavy rain, cold wave, arid and is
Can be dead, be a kind of slow-growing, the low-down orchid family of self-reproduction ability is possessed a person plant, due to the stem of noble dendrobium have it is huge medicinal
And health value, cause long-term predatory excavation, along with living environment is more and more severe, cause wild resource seriously exhausted.
Nowadays stem of noble dendrobium reproduction technique has seed to sprout, cuttage, division propagation, and tissue cultures, wherein seed sprouting germination rate are extremely low, skewer
Slotting, division propagation reproduction speed is also very slow.Tissue rapid propagation technology is a kind of method for effectively improving seeling industry efficiency, with group
Knitting culture and carry out the cultivation of the stem of noble dendrobium can greatly improve the reproductive efficiency of the stem of noble dendrobium.
Production at present is upper to be carried out expanding numerous using callus induction mostly, but is debugged on hormone combination,
However, hormone combination debugging is difficult to meet optimum condition needed for the stem of noble dendrobium, so as to cause bud ratio relatively low.
The content of the invention
The main object of the present invention is to propose a kind of method for growing dendrobium seedlings, it is intended to improve the bud ratio of growing dendrobium seedlings.
To achieve the above object, the present invention proposes a kind of method for growing dendrobium seedlings, comprises the following steps：
A, the stem of noble dendrobium stem section of belt segment is cultivated in the medium, induce axillary bud；
B, when axillary bud growth is highly more than 1cm, by bud point excision, induce Multiple Buds；
C, when the growth of all living creatures' bud reaches 1cm~2cm, cut Multiple Buds, Multiple Buds are rised in value and culture of rootage, and will
The former stem section for having cut Multiple Buds continues to cultivate in the medium, evoked callus；
D, after Callus formation, callus is cut and is transferred in culture medium and continues to cultivate, induce Multiple Buds；
E, Multiple Buds are transferred to Multiplying culture in culture medium；
F, when Multiple Buds growing height reaches 2cm~3cm, being transferred in culture medium carries out culture of rootage.
Preferably, above-mentioned steps b also includes：When bud point is cut off, the bud point of reservation 0.1cm~0.5cm is in stem section.
Preferably, above-mentioned steps b also includes：After bud point is cut off, rip cutting is carried out on the tangent plane that the bud to retaining is selected.
Preferably, the stem section chosen in step a is the stem section near root of the stem of noble dendrobium.
Preferably, above-mentioned steps a also includes：Before medium culture, stem section is carried out disinfection, sterilization processing.
Preferably, the formula to culture medium in step a is：MS, NAA：0.4mg/L~0.6mg/L, carbon dust：0.2~
1.0g/L, 6-BA：1.0mg/L~2.0mg/L, KT：0.2mg/L~0.4mg/L, banana：10g/L~25g/L.
Preferably, the formula to culture medium in step c is：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：
40g/L~60g/L, carbon dust 0.2g/L~1.0g/L, NAA：0.3mg/L~0.4mg/L, 6-BA：0mg/L~2.0mg/L, KT：
0.1mg/L~0.3mg/L, carbon dust：0.2~1.0g/L.
Preferably, the formula of culture medium is in step d：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：
40g/L~60g/L, carbon dust 0.2g/L~1.0g/L, NAA：1.0mg/L~1.2mg/L, 6-BA：1.5mg/L~1.7mg/L.
Preferably, the formula of culture medium is in step e：1/2MS culture mediums, banana：100g/L~120g/L, carbon dust
0.2g/L~1.0g/L, NAA：0.5mg/L~1.5mg/L, IBA：0.5mg/L~1.0mg/L.
Technical scheme cuts to break apical dominance by bud point, promotes to sprout more Multiple Buds, shortens
Cultivation cycle, and the cutting for passing through Multiple Buds, to promote the formation of evoked callus, carry out expanding numerous using callus, can
Numerous quantity is expanded with effective increase, so as to produce the stem of noble dendrobium seedling of a large amount of high-quality.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
The accompanying drawing to be used needed for having technology description is briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those of ordinary skill in the art, on the premise of not paying creative work, can be with
Structure according to these accompanying drawings obtains other accompanying drawings.
Fig. 1 is the tiller number after Dendrobidium huoshanness Multiplying culture about 60d；
Fig. 2 is the growth value after Dendrobidium huoshanness Multiplying culture 60d；
Fig. 3 is Dendrobidium huoshanness by the culture of rootage plant height figure after potato treatment；
Fig. 4 is that Dendrobidium huoshanness is slightly schemed by the culture of rootage stem after potato treatment；
Fig. 5 is Dendrobidium huoshanness by the culture of rootage radical figure after potato treatment；
Fig. 6 is that Dendrobidium huoshanness is slightly schemed by the culture of rootage root after potato treatment；
Fig. 7 is Dendrobidium huoshanness by the culture of rootage plant height figure after banana treatment；
Fig. 8 is that Dendrobidium huoshanness is slightly schemed by the culture of rootage stem after banana treatment；
Fig. 9 is Dendrobidium huoshanness by the culture of rootage radical figure after banana treatment；
Figure 10 is that Dendrobidium huoshanness is slightly schemed by the culture of rootage root after banana treatment.
The realization of the object of the invention, functional characteristics and advantage will be described further referring to the drawings in conjunction with the embodiments.
Specific embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the invention, rather than whole embodiments.Base
Embodiment in the present invention, those of ordinary skill in the art obtained under the premise of creative work is not made it is all its
His embodiment, belongs to the scope of protection of the invention.
If it is to be appreciated that related in the embodiment of the present invention directionality indicate (such as up, down, left, right, before and after ...),
Then directionality indicate to be only used for explain relative position relation between each part under a certain particular pose (as shown in drawings),
Motion conditions etc., if the particular pose changes, directionality indicates also correspondingly to change therewith.
If in addition, relating to the description of " first ", " second " etc. in the embodiment of the present invention, being somebody's turn to do " first ", " second " etc.
Description be only used for describing purpose, and it is not intended that indicating or implying its relative importance or implicit indicate indicated skill
The quantity of art feature.Thus, " first " is defined, at least one spy can be expressed or be implicitly included to the feature of " second "
Levy.In addition, the technical scheme between each embodiment can be combined with each other, but must be with those of ordinary skill in the art's energy
Based on enough realizations, when the combination appearance of technical scheme is conflicting or cannot realize it will be understood that the knot of this technical scheme
Conjunction does not exist, not within the protection domain of application claims yet.
The present invention proposes a kind of method for growing dendrobium seedlings.
In embodiments of the present invention, method for growing dendrobium seedlings is comprised the following steps：
A, the stem of noble dendrobium stem section of belt segment is cultivated in the medium, induce axillary bud.
Selection is sturdy, and gum level is high, the Dendrobidium huoshanness plant of no disease and pests harm, removes blade, and liquid detergent is cleaned up, and flows
Water is rinsed about 1 hour, and distilled water is cleaned up, and is wiped over totally with 70% alcohol on superclean bench, removal leaf sheath, and will
The fresh bar of the stem of noble dendrobium is cut into the stem-segment with node of about 1.5cm.Then 5min-8min is soaked by about 0.1% mercuric chloride, then it is left with 10%
Right liquor natrii hypochloritis sterilization 8min, with aseptic water washing 5-6 times.Stem section is finally kept flat culture in being inoculated in culture medium big
About 15 days.
B, when axillary bud growth is highly more than 1cm, by bud point excision, induce Multiple Buds.
In step a, stem section was cultivated by 15 days, and the milk green bud point of about 1cm high is grown at node, was now cut
Bud point.Concrete operations are as follows：
1. bud point is cut off with blade, that is, removes terminal bud, remove apical dominance；Then it is inoculated into culture medium
In.It should be noted that in cutting, it is impossible to cut bud point completely, the bud point that should retain about 0.1cm~0.5cm or so exists
In stem section, such as including 0.3cm.In order to verify that bud point retains the relation of length and the quantity of Multiple Buds, experiment number table is as follows：
| Bud point retains length (cm) | 30 stem sections, Multiple Buds par after 15d |
| 0 | 0.2 |
| 0.1 | 4.6 |
| 0.2 | 5.6 |
| 0.3 | 5.5 |
| 0.4 | 5.3 |
| 0.5 | 4.9 |
| 0.6 | 3.5 |
| 0.7 | 2.7 |
Thus, it can be seen that, when bud point retains length in 0.1~0.5cm, the quantity of Multiple Buds is most.When bud point whole quilt
After excision, almost without Multiple Buds.When bud point retains length more than 0.6cm, it is clear that the negligible amounts of Multiple Buds, inducing effect
It is unobvious.
2. partial application, is indulged on the tangent plane that the bud for retaining is selected with blade, to increase otch area, callus success is improved
Rate.Then continue to be put into and cultivate about 25 days in former culture medium.
C, whne all living creatures' bud growth reach 1cm~2cm when, cut Multiple Buds, and will cut Multiple Buds former stem section continuation
Cultivate in the medium, evoked callus.
Cultivated by 25 days or so, stem section incision grows 4-6 Multiple Buds, and about 1-2cm or so high will now cut clump
Sprout.Behaviour is as follows for step：
Adventitious buds proliferation：Multiple Buds under cutting are transferred in culture medium and carry out culture 60 days.Pass through culture medium after 60 days
Carry out culture of rootage.
Callus induction：The former induction stem section that Multiple Buds will have been cut continues to be put into evoked callus in culture medium.
D, after Callus formation, callus is cut and is transferred in culture medium and continues to cultivate, induce Multiple Buds.
By the culture of about 30 days, stem section otch produced callus, part callus it is differentiated go out Multiple Buds,
An average stem section produces budlet 6-8 or so that grows thickly, and callus is cut into by knife in fritter continuation culture medium.
Again by the culture of about 30 days, stem section Multiple Buds quantity is increased sharply, and callus all sprout by differentiation, becomes clump
Sprout cluster, Multiple Buds are now cut out switching to carry out Multiplying culture by height probably 1cm or so, and callus can continue to be transferred to
Culture medium carries out inducing clumping bud culture.
E, Multiple Buds are transferred to Multiplying culture in culture medium.
In order to improve quantity of sprouting, all living creatures' bud in previous step is transferred in culture medium carries out Multiplying culture, to obtain
More all living creatures' buds.
F, when Multiple Buds growing height reaches 2cm~3cm, being transferred in culture medium carries out culture of rootage.
By the culture of step e, stem of noble dendrobium seedling is sufficiently robust after cultivating about 60 days, height probably 2-3cm or so, can carry out
Culture of rootage.The stem of noble dendrobium seedling that will be turned out is taken root in being transferred to culture medium.
G, emerge.
After culture culture of rootage about 75 days, stem of noble dendrobium seedling grows root system to stem of noble dendrobium seedling, and stem of noble dendrobium root is sent out up to 5cm or so and root
Green, stem of noble dendrobium seedling reaches more than 7-8cm, grows blade more than 5, and leaf color is dark green, you can remove outdoor acclimatization and transplantses.
Technical scheme cuts to break apical dominance by bud point, promotes to sprout more Multiple Buds, shortens
Cultivation cycle, and the cutting for passing through Multiple Buds, to promote the formation of evoked callus, carry out expanding numerous using callus, can
Numerous quantity is expanded with effective increase, so as to produce the stem of noble dendrobium seedling of a large amount of high-quality.
Stem of noble dendrobium bar upper grown cellulose content is higher, in theory, when stem section is chosen, top should be preferentially chosen, with training
More Multiple Buds are grown when foster.In order to verify influence of the stem section of each and every one position of stem of noble dendrobium bar to Multiple Buds quantity, choose suddenly
The mountain stem of noble dendrobium is material, and experiment number table is as follows：
As can be seen here, it is not that the Multiple Buds quantity that top stem section is produced is most right, conversely, leaning on for Dendrobidium huoshanness
The Multiple Buds that nearly root stem section is produced are more, and this is on the contrary with the cognition of technical staff.
Therefore, when stem of noble dendrobium stem section is chosen, preferably adjacent to the stem section of root.
In the description below, by with the one kind in the stem of noble dendrobium, it is illustrated as a example by the cultivation of Dendrobidium huoshanness.
In above-mentioned steps a, although the culture medium for cultivating stem of noble dendrobium stem section can have various, but, in the present embodiment, lead to
Experiment is crossed, preferably M1 culture medium prescriptions are drawn.
Experiment is as follows：
With MS as minimal medium, addition banana 10g/L~25g/L is (for the stem of noble dendrobium provides various nutrients, such as amino
Acid and trace element etc.), NAA, carbon dust, KT, 6-BA, 9 kinds of culture mediums of hormon proportioning of configuration, specific hormone prescription such as table
One：
The stem section of table one induces Multiple Buds culture medium prescription table
Statistical observation can obtain (table two) after 40d, and from the point of view of 9 kinds of culture mediums of inoculation, No. 9 culture mediums are that hormone combination is 6-
The axillary bud deriving rate highest of BA2mg/L+KT0.4mg/L, the axillary bud of induction reaches 102, and the bud-leaf color of induction is dark green, sturdy,
Height highest.Next to that No. 6 culture mediums are that hormone combination is 6-BA1mg/L+KT0.4mg/L, the axillary bud number of induction is 90, is lured
The bud-leaf color led is green, more sturdy.It is again that 5 i.e. hormone combination for the treatment of is 6-BA1mg/L+KT0.2mg/L.Worst is 1,2,
No. 7 culture mediums, do not sprout axillary bud substantially, and only minority sprouts axillary buds and sprouting axillary bud number is less, and axillary bud is thin and delicate, and growing way is weaker,
Only only a few has sprouting phenomenon and robustness is weaker, is not suitable for carrying out the switching of next step.Therefore 1,2, No. 7 culture medium discomforts
Cooperate to induce Multiple Buds culture medium for Dendrobidium huoshanness stem section.
The stem section of table two induces Multiple Buds 60d statistical forms
++++very healthy and strong；+++ it is healthy and strong；++ it is general healthy and strong；+ weak
Stem-segment with node needs to sprout comparatively ideal axillary bud under the collective effect of the basic element of cell division and auxin.
In certain limit, the basic element of cell division KT and 6-BA and the NAA of low concentration of higher concentration cooperate with the induction for being conducive to bud, hormone
Excessive concentration or the too low induction for being all unfavorable for axillary bud are sprouted, in the case that 6-BA concentration is certain, KT stems higher in certain limit
Section induction axillary bud number is more.Consider, Dendrobidium huoshanness stem section induces the relatively suitable culture medium of Multiple Buds for minimal medium is:
MS, NAA：0.4mg/L~0.6mg/L, 6-BA：1.0mg/L~2.0mg/L, KT：0.2mg/L~0.4mg/L, banana：10g/L
~25g/L.Now Multiple Buds are cut carries out Multiplying culture, and remaining stem section carries out the induction of callus.Need herein
It is bright, the easy brown stain in incubation due to stem section, because amino-compound such as protein in culture medium contained by it,
Amino acid and aldehyde, ketone etc. meet with reduced sugar, and the phenomenon by series reaction generation brown polymer is referred to as browning reaction, and
Brown stain can cause stem section to be germinateed.A certain amount of carbon dust is added, brown stain can be effectively prevented, so that improve from the bud ratio sprouted,
This, carbon dust addition in the range of 0.2~1.0g/L, such as 0.3g/L, 0.5g/L or 0.8g/L.It is following in order to prevent brown stain
Carbon dust will be in the lump added in other embodiments.Therefore, the minimal medium M1 of Dendrobidium huoshanness is：MS, NAA：0.4mg/L~
0.6mg/L, 6-BA：1.0mg/L~2.0mg/L, KT：0.2mg/L~0.4mg/L, banana：10g/L~25g/L, carbon dust：
0.2g/L~1.0g/L.
In above-mentioned steps c, by orthogonal design, preferably M3 culture medium prescriptions are drawn.
Design Three factors-levels orthogonal experiment, each factor meter is table three, and orthogonal design table is table four, and minimal medium is
Spend precious No. 4：1.0~2.0g/L, banana：40g/L~60g/L, carbon dust 0.2g/L~1.0g/L.The first step is tested into remaining stem
Section is inoculated in 9 treatment in orthogonal design table, each 5 bottles for the treatment of inoculation, and 5 stem sections of every bottle of inoculation observe callus during 30d
Tissue induction number, observes callus induction bud number after 60d.
The factor level table of table three
The callus induction orthogonal design table of table four
Each observation can obtain each treatment after 30d generation callus, continues to find that callus is equal after cultivating 60d
Adventitious bud, experimental result such as table five are produced.
Being analyzed by orthogonal experiment to obtain, and the suitable level of individual factor is, NAA：0.3mg/L, 6-BA：1mg/L, KT：
0.3mg/L, and individual factor influence size is NAA>KT>6-BA.
Therefore the M3 culture medium prescriptions of suitable stem section evoked callus are：1/2MS culture mediums, spend precious No. 4：1.0~
2.0g/L, banana：40g/L~60g/L, carbon dust 0.2g/L~1.0g/L, NAA：0.3mg/L or so, 6-BA：0mg/L~
2.0mg/L, KT：0.1mg/L~0.3mg/L, carbon dust：0.2~1.0g/L.
The callus induction experimental result of table five
The callus induction experimental result of table six
By table six, it can be deduced that, NAA is in 0.3mg/L~0.4mg/L, and callus induction number and adventitious bud are average
Induction number highest, therefore, M3 culture medium prescriptions are：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：40g/L~
60g/L, carbon dust 0.2g/L~1.0g/L, NAA：0.3mg/L~0.4mg/L, 6-BA：0mg/L~2.0mg/L, KT：0.1mg/L
~0.3mg/L, carbon dust：0.2~1.0g/L.
In above-mentioned steps e, by orthogonal design, preferably M2 culture medium prescriptions are drawn.
Minimal medium is：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：40g/L~60g/L, carbon dust
0.2g/L~1.0g/L.Add NAA, 6-BA, 9 proliferated culture mediums of hormon proportioning of configuration, culture medium of various concentrations
Formula such as table five, will be highly consistent, and the propagation seedling of about 1cm is inoculated in this 9 culture mediums of different disposal, and each treatment connects
Kinds 20 bottles, 5 Dendrobidium huoshanness propagation seedlings of every bottle of inoculation, after about 60d statistical observation and record seedling tiller number, growth value,
Color, growth potential.
Growth potential：+ growing way is weaker, and color is more yellow, and seedling is short or very thin；++ growing way is general, and color is yellowish green, and seedling is too high or fibre
Carefully；+++ growing way is preferable, colors green；+++ preferably, color is dark green, and seedling is sturdy for+growing way.
The Dendrobidium huoshanness of table seven increment culture 60d statistical forms
Statistical observation can be obtained after 60d, and from the point of view of tiller number (reference picture 1), treatment 5 preferably, next to that processing 2, processes 4,
Treatment 3,9 tiller numbers for the treatment of are worst.From the point of view of growth value (reference picture 2), treatment 5 preferably, next to that treatment 4, processes 2, is processed
1, same treatment 9 is worst.From the point of view of growth potential, color, preferably, color is dark green, and seedling is sturdy for treatment 4 and treatment 5, processes 1 and place
Reason 2 is taken second place, and preferably, colors green, treatment 7 and treatment 9 are worst, and growing way is most weak for growing way.
Comprehensive analysis can be obtained, and treatment 5,4 optimums for the treatment of do Dendrobidium huoshanness Multiplying culture culture medium.Wherein treatment 5 is best
That is NAA1mg/L+6-BA1.5mg/L combination optimums do Dendrobidium huoshanness Multiplying culture culture medium.Treatment 4 is not added with its points of 6-BA
Tiller relatively process 5 and treatment it is 2 poor, but seedling growing way preferably, and experiment find that the seedling for processing 4 need not carry out strong plantlets and rootage can be real
Existing forming seedling through one step culture, had not only saved the means of production but also had shortened the time, and drawback is to emerge quality slightly poorer to the kind for carrying out strong plantlets and rootage
Seedling, and quantity of emerging is slightly less than the rooted seedling of a switching generation.Experiment finds that the certain density 6-BA and NAA of addition can promote Huoshan
Stem of noble dendrobium proliferation-inducing, NAA concentration is constant, within the specific limits, with the rising of 6-BA concentration its proliferation-inducing effect also gradually
Become strong, it is most strong that experiment can add 6-BA1.5mg/L inducing effects, but concentration, when being higher than 6-BA1.5mg/L, inducing effect is on the contrary
Die down, illustrating the 6-BA of high concentration can suppress Dendrobidium huoshanness Multiplying culture.But in the case that 6-BA concentration is constant, addition is certain dense
Spend gradient NAA, its cultivation effect be also as the rising of NAA concentration is first strengthened weakening afterwards, therefore high concentration NAA to Huoshan
The cultivation effect of the stem of noble dendrobium is unhelpful.
The Dendrobidium huoshanness of table eight increment culture 60d statistical forms
Can be obtained by table eight, NAA：1.0mg/L~1.2mg/L, 6-BA：During 1.5mg/L~1.7mg/L, Dendrobidium huoshanness
Growth formula and color are all best.
Comprehensive analysis can be obtained, and the culture medium of optimum Dendrobidium huoshanness Multiplying culture is 1/2MS culture mediums+spend No. 4 1.5g/ of treasured
L+ banana 50g/L+ carbon dusts 0.5g/L+NAA1mg/L+6-BA1.5mg/L.
M2 culture medium prescriptions：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：40g/L~60g/L, carbon dust
0.2g/L~1.0g/L, NAA：1.0mg/L~1.2mg/L, 6-BA：1.5mg/L~1.7mg/L.
In above-mentioned steps f, by orthogonal design, preferably M4 culture medium prescriptions are drawn.
Minimal medium is 1/2MS culture mediums, carbon dust 0.2g/L~1.0g/L, and add NAA, IBA of various concentrations,
9 root medias of hormon proportioning of configuration, additive selects banana and potato respectively, and concentration is respectively 100g/L.Training
Based formulas such as table five is supported, will be highly consistent, the propagation seedling of about 2.5cm is inoculated in this 9 culture mediums of different disposal, each
20 bottles for the treatment of inoculation, every bottle of inoculation 5 Dendrobidium huoshanness propagation seedlings, 75d statistical observations once and record seedling growth potential, stalwartness
Degree, height of seedling, stem are thick, radical, root are thick.Condition of culture is about 25 DEG C of temperature, about 2000lx10 hour of illumination.
The Dendrobidium huoshanness root-promoting hormone of table nine matches table
Test tube seedling is taken out after each treatment culture about 75d, is cleaned, measure plant height, stem is thick, radical, root are thick, and comparative analysis is such as
Fig. 3 to Figure 10.Analysis result is as follows：
Banana processes plant height (reference picture 7)：After addition 100g/L banana cultures 40d, 8 are processed, hormone combination is
NAA1.5mg/L+IBA0.5mg/L plant heights are up to 8.2cm, secondly for 1 hormone combination for the treatment of is NAA0.5mg/L+IBA0mg/L
Plant height is 6.5cm, and treatment 7 is minimum, and hormone combination is only 4.8cm for NAA1.5mg/L+IBA0mg/L plant heights.The NAA of high concentration
(1.5mg/L) coordinates the certain density IBA (0.5mg/L) to be effectively promoted increasing for Dendrobidium huoshanness rooted seedling.
Banana treatment stem is thick (reference picture 8)：After addition 100g/L banana cultures 40d, stem of Dendrobium slightly averagely reaches 0.34cm,
Wherein treatment 8 is that hormone combination is treatment 2 i.e. hormone for NAA1.5mg/L+IBA0.5mg/L stems are slightly 0.39cm to the maximum, secondly
Match be NAA0.5mg/L+IBA0.5mg/L, treatment 5 i.e. hormone combination for secondly NAA1mg/L+IBA0.5mg/L stems are slightly
0.38cm, treatment 6 i.e. hormone combination is that NAA1mg/L+IBA1mg/L stems are slightly minimum, only 0.28cm.Individually add finite concentration
The NAA growth results thick to stem it is unobvious, and the NAA (1.5mg/L) for adding higher concentration coordinates certain density IBA
(0.5mg/L) can be effectively promoted the cross growth of Dendrobidium huoshanness stem, and enhancing stem is thick.
Banana treatment radical, root are thick (reference picture 9 and Figure 10)：Radical substantially increases after addition 100g/L banana cultures 40d,
Each treatment radical is more, and mean elements reaches 8, wherein treatment 1 i.e. hormone combination is NAA0.5mg/L+IBA0mg/L roots
Number is maximum, is 10, and it is that NAA0.5mg/L+IBA1mg/L, the i.e. hormone combinations for the treatment of 8 are to process 3 i.e. hormone combination
NAA1.5mg/L+IBA0.5mg/L radicals are more, and mean elements reaches more than 8.5.Therefore banana can be effectively promoted Huoshan
Stem of noble dendrobium root of hair.And banana processes its root and slightly averagely reaches 1.1mm, wherein treatment 3 i.e. hormone combination is NAA0.5mg/L+
IBA1mg/L roots are slightly 0.14cm to the maximum, and process 6 i.e. hormone combination NAA1mg/L+IBA1mg/L and treatment 1 i.e. hormone combination
Secondly NAA0.5mg/L+IBA0mg/L roots are slightly 0.12cm.The NAA cultures of simple plus low concentration can be effectively promoted the transverse direction of root
Growth.
In the Dendrobidium huoshanness culture of rootage culture medium of addition banana treatment, comprehensive rooted seedling plant height, stem are thick, radical, root are thick
These index analysis can be obtained, and the hormone combination for being adapted to Dendrobidium huoshanness strong plantlets and rootage is NAA1.5mg/L+IBA0.5mg/L.
Potato processes plant height (reference picture 3)：After addition 100g/L potato cultures 40d, 7 are processed, hormone combination is
NAA1.5mg/L+IBA0mg/L plant heights are up to 6.7cm, secondly for 9 hormone combinations for the treatment of are NAA1.5mg/L+IBA1mg/L plants
A height of 6.2cm, 1 hormone combination for the treatment of is that NAA0.5mg/L+IBA0mg/L plant heights are 5.8cm, and treatment 2 is minimum, and hormone combination is
NAA0.5mg/L+IBA0.5mg/L plant heights are only 5.34cm.The NAA (1.5mg/L) of high concentration can be effectively promoted Dendrobidium huoshanness
Rooted seedling increases.
Potato treatment stem is thick (reference picture 4)：After addition 100g/L potato cultures 40d, each treatment stem of Dendrobium rough error is different not
Greatly, more than 0.3cm is reached, treatment 1 i.e. hormone combination is that the thick relative maximum of NAA0.5mg/L+IBA0mg/L stems is 0.4cm.Place
Reason 2 is that hormone combination is：NAA0.5mg/L+IBA0.5mg/L, treatment 4 i.e. hormone combination are cultivated for NAA1mg/L+IBA0mg/L
It is 0.37cm that stem is slightly relatively fewer, and it is the thick minimum 0.29cm of NAA1mg/L+IBA0.5mg/L stems to process 5 i.e. hormone combination,
Maximum is only 1mm in minimum value difference, and difference is little.Therefore the hormon of addition potato treatment is thick with stem of Dendrobium is compared
Increase impact effect little.
Potato treatment radical, root are thick (reference picture 5 and Fig. 6)：After addition 100g/L potato cultures 40d, each treatment stem of noble dendrobium
Radical and root rough error are different less.Radical aspect treatment 9 i.e. hormone combination is 7.8 to the maximum for NAA1.5mg/L+IBA1mg/L radicals,
Treatment 6 i.e. hormone combination is NAA1mg/L+IBA1mg/L and the i.e. hormone combination for the treatment of 1 is：NAA0.5mg/L+IBA0mg/L radicals
Take second place, be followed successively by 5.9 and 5.1.The thick aspect treatment 1 of root i.e. hormone combination is NAA0.5mg/L+IBA0mg/L and treatment 8 i.e. hormone
Match as NAA1.5mg/L+IBA0.5mg/L roots are slightly 0.14cm to the maximum, treatment 4 i.e. hormone combination is NAA1mg/L+IBA0mg/
L roots are thick to small only 0.08cm.
In the Dendrobidium huoshanness culture of rootage culture medium of addition potato treatment, comprehensive rooted seedling plant height, stem are thick, radical, root are thick
These index analysis can be obtained, and the hormone combination for being adapted to Dendrobidium huoshanness strong plantlets and rootage is NAA0.5mg/L+IBA0mg/L.
Comprehensive analysis can be obtained：Addition banana treatment Dendrobidium huoshanness rooted seedling growing way is generally handled well than addition potato, is added
Banana treatment group stem of noble dendrobium plant height and radical are substantially handled well than addition potato, and stem is thick and root rough error is not different notable, and addition is fragrant
Any of several broadleaf plants treatment stem of noble dendrobium seedling color is dark green, and addition potato treatment stem of noble dendrobium seedling color is light green.Comprehensive analysis can be obtained, and addition banana 100g/L is more
Suitable Dendrobidium huoshanness culture of rootage.
As can be seen from Table 10, either from plant height, radical or root it is thick from, banana concentration value be 100g/L~
During 120g/L, stem of noble dendrobium root growth is best.
The Dendrobidium huoshanness root-promoting hormone of table ten matches table
It can thus be concluded that, suitable Dendrobidium huoshanness strong plantlets and rootage M4 culture mediums are：1/2MS culture mediums, banana：100g/L~
120g/L, carbon dust 0.2g/L~1.0g/L, NAA：0.5mg/L~1.5mg/L, IBA：0.5mg/L~1.0mg/L.
The preferred embodiments of the present invention are the foregoing is only, the scope of the claims of the invention is not thereby limited, it is every at this
Under the inventive concept of invention, the equivalent structure transformation made using description of the invention and accompanying drawing content, or directly/use indirectly
It is included in scope of patent protection of the invention in other related technical fields.
Claims (10)
1. a kind of method for growing dendrobium seedlings, it is characterised in that comprise the following steps：
A, the stem of noble dendrobium stem section of belt segment is cultivated in the medium, induce axillary bud；
B, when axillary bud growth is highly more than 1cm, by bud point excision, induce Multiple Buds；
C, when the growth of all living creatures' bud reaches 1cm~2cm, cut Multiple Buds, Multiple Buds are rised in value and culture of rootage, and will cutting
The former stem section of complete Multiple Buds is cultivated in being put into culture medium, evoked callus；
D, after Callus formation, callus is cut and is transferred in culture medium and continues to cultivate, induce Multiple Buds；
E, Multiple Buds are transferred to Multiplying culture in culture medium；
F, when Multiple Buds growing height reaches 2cm~3cm, being transferred in culture medium carries out culture of rootage.
2. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that above-mentioned steps b also includes：When bud point is cut off,
The bud point of reservation 0.1cm~0.5cm is in stem section.
3. method for growing dendrobium seedlings as claimed in claim 2, it is characterised in that above-mentioned steps b also includes：After bud point is cut off,
Rip cutting is carried out on the tangent plane selected to the bud for retaining.
4. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that the stem section chosen in step a is close to for the stem of noble dendrobium
The stem section of root.
5. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that above-mentioned steps a also includes：Medium culture it
Before, stem section is carried out disinfection, sterilization processing.
6. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that the formula to culture medium in step a is：MS,
NAA：0.4mg/L~0.6mg/L, 6-BA：1.0mg/L~2.0mg/L, KT：0.2mg/L~0.4mg/L, banana：10g/L~
25g/L。
7. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that to the propagation institute of Multiple Buds in step c and step e
The formula of the culture medium for using for：1/2MS culture mediums, spend precious No. 4：1.0~2.0g/L, banana：40g/L~60g/L, carbon dust
0.2g/L~1.0g/L, NAA：0.3mg/L~0.4mg/L, 6-BA：0mg/L~2.0mg/L, KT：0.1mg/L~0.3mg/L.
8. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that the formula of culture medium is in step d：1/2MS is trained
Base is supported, spends precious No. 4：1.0~2.0g/L, banana：40g/L~60g/L, NAA：1.0mg/L~1.2mg/L, 6-BA：1.5mg/L
~1.7mg/L.
9. method for growing dendrobium seedlings as claimed in claim 1, it is characterised in that the formula of culture medium is in step e：1/2MS is trained
Support base, banana：100g/L~120g/L, NAA：0.5mg/L~1.5mg/L, IBA：0.5mg/L~1.0mg/L.
10. the method for growing dendrobium seedlings as described in claim 6 to 9 any one, it is characterised in that be also added with culture medium
The carbon dust of 0.2~1.0g/L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646678A (en) * | 2017-09-28 | 2018-02-02 | 石阡县龙腾农牧农民专业合作社 | A kind of method for growing dendrobium seedlings |
| CN109984035A (en) * | 2019-03-22 | 2019-07-09 | 扬州雅典娜园艺科技开发有限公司 | A method of establishing HERBA DENDROBII regenerating system |
| CN111066656A (en) * | 2020-01-16 | 2020-04-28 | 贵州师范大学 | Culture medium group and method for dendrobium officinale tissue culture |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101258835A (en) * | 2008-04-23 | 2008-09-10 | 昆明理工大学 | Fast reproducing method for high quality seedling of dendrobium officinale |
| CN104322375A (en) * | 2014-11-21 | 2015-02-04 | 广西中医药大学 | Method for rapidly propagating dendrobium chrysotoxumLindl. seeds by tissue culture |
-
2016
- 2016-11-16 CN CN201611027267.5A patent/CN106718875B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101258835A (en) * | 2008-04-23 | 2008-09-10 | 昆明理工大学 | Fast reproducing method for high quality seedling of dendrobium officinale |
| CN104322375A (en) * | 2014-11-21 | 2015-02-04 | 广西中医药大学 | Method for rapidly propagating dendrobium chrysotoxumLindl. seeds by tissue culture |
Non-Patent Citations (2)
| Title |
|---|
| STADWELSON DOHLING等: "Multiple shoot induction from axillary bud cultures of the medicinal orchid ，Dendrobium longicornu", 《AOB PLANTS》 * |
| 彭颖等: "铁皮石斛组织培养技术", 《农业工程》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646678A (en) * | 2017-09-28 | 2018-02-02 | 石阡县龙腾农牧农民专业合作社 | A kind of method for growing dendrobium seedlings |
| CN109984035A (en) * | 2019-03-22 | 2019-07-09 | 扬州雅典娜园艺科技开发有限公司 | A method of establishing HERBA DENDROBII regenerating system |
| CN111066656A (en) * | 2020-01-16 | 2020-04-28 | 贵州师范大学 | Culture medium group and method for dendrobium officinale tissue culture |
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